LOCUS       sequence-172879-        7781 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encrypted AND gate, decryption step 2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7781)
  TITLE     Tim Lu lab plasmids
REFERENCE   2  (bases 1 to 7781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pOP447
FEATURES             Location/Qualifiers
     source          1..7781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(519..1469)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1517..1739
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2243..2337)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2367..3158,3159..3227))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2990..3009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3228..3332)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    3436..3469
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    3619..3668
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      complement(3682..3768)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3774..4652)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        complement(join(4866..4891,4892..4897,4898..4914,4915..4920,
                     4921..4939))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4896..4914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4921..4939
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(4953..5039)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5045..6127)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6089..6108
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6254..6311
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    6314..6330
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             6532..6561
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    6739..6758
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    6772..6788
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_RNA        6855..6904
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(6988..7016)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7611..7631
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      complement(7681..7775)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc
       61 gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag aattgccatg
      121 attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa
      181 gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca
      241 ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg
      301 ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttgaatgcac
      361 caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata
      421 tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg tttgttagtc
      481 ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc
      541 agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga
      601 tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt
      661 gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga atctgatgta
      721 atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg
      781 agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc
      841 ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa
      901 acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta
      961 aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt
     1021 aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga
     1081 ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact
     1141 aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct
     1201 ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct
     1261 aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg
     1321 aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca
     1381 cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca
     1441 tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca
     1501 ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg
     1561 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
     1621 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
     1681 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
     1741 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
     1801 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
     1861 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     1921 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     1981 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     2041 agggcgtttt atggtgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     2101 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     2161 tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc
     2221 tagtatcgat actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2281 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2341 gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3361 gtccttacgt gccgatcaac gtctcatttt cgccagatat cgacgtcaaa gattctagga
     3421 tctaaccgta tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actcccacag
     3481 ctaacaccac gtcgtcccta tctgctgccc taggtctatg agtggttgct ggataacttt
     3541 acgggcatgc ataaggctcg tataatatat tcagggagac cacaacggtt tccctctaca
     3601 aataattttg tttaactttc ggccggcttg tcgacgacgg cggtctccgt cgtcaggatc
     3661 atccgggctc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     3721 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gccttatgac
     3781 aacttgacgg ctacatcatt cactttttct tcacaaccgg cacgaaactc gctcgggctg
     3841 gccccggtgc attttttaaa tactcgcgag aaatagagtt gatcgtcaaa accaacattg
     3901 cgaccgacgg tggcgatagg catccgggta gtgctcaaaa gcagcttcgc ctggctgata
     3961 cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac
     4021 agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct
     4081 gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggcggatgg
     4141 agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc
     4201 agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg
     4261 ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaaac ccgtattggc aaatattgac
     4321 ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac
     4381 cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa
     4441 atatcacccg gtcggcagac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga
     4501 atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga
     4561 taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcgttaaa cgagtatccc
     4621 ggcagcaggg gatcattttg cgcttcagcc atgctagctc tcgtccctag cttcgccgaa
     4681 ggcgttaaac aaaattattt gtagaggggg tttcgtcatc actgactcat cagagttgaa
     4741 agcacatcaa ctgacccctg tttgtaaagg agactgataa tggcatgtac aactatacta
     4801 gttgtagtgc ctaaataatg cttcccccaa ctgagagaac tcaaaggtta ccccagttgg
     4861 ggcacggtca gtgcgtcctg ctgatgtgct cagtatctct atcactgata gggatgtcaa
     4921 tctctatcac tgatagggat ctagggcggc ggatttgtcc tactcaggag agcgttcacc
     4981 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     5041 tgccttactg cccgctttcc agtcgggaaa cctgtcgtgc cagttgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggagccag ggtggttttt cttttcacca
     5161 gtgagacggg caacagttga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     5221 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     5281 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc
     5341 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     5401 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     5461 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     5521 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     5581 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     5641 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     5701 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     5761 tcagcccact gacgcgctgc gcgagaagat tgtgaaccgc cgctttacag gcttcgacgc
     5821 cgcttcgttc taccatcgac accaccacgc tggcacccag ctggtcagca cgagatttga
     5881 tagcagcaac gatctgagac ggagcgtgca gagccagaga agaggtggca acgccaatca
     5941 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     6001 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     6061 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtac agcgttactg
     6121 gtttcatgtg cactacctcc ttggttgtct atgctatgct gatctacaac tggcatgctt
     6181 aaacaaaatt atttgtagag gtagtttcgc cttcacaggc tcatcagacc ggcaagcaca
     6241 gccggtgact actgtcgggg tttgtaccgt acaccactga gaccgcggtg gttgaccaga
     6301 caaaccacga caattgtgag cggataacaa tttcacacag ggccctcgga caccgaggag
     6361 aatgtcaaga ggcgaacaca caacgtcttg gagcgccaga ggaggaacga gctaaaacgg
     6421 agcttttttg ccctgcgtga ccagatcccg gagttggaaa acaatgaaaa ggcccccaag
     6481 gtagttatcc ttaaaaaagc cacagcatac atcctgtccg tccaagcaga ggagcaaaag
     6541 ctcatttctg aagaggactt gttgcggaaa cgacgagaac agttgaaaca caaacttgaa
     6601 cagctacgga actcttgtgc gtaaggaaaa gtaaggaaaa cgattccttc taacagaaat
     6661 gtcctgagca atcacctatg aactgtcgac tcgagcatag catttttatc cataagatta
     6721 gcggatccta agctttacaa ttgtgagcgc tcacaattat gatagattca attgtgagcg
     6781 gataacaatt tcacacagca aaaaaagcaa aaagcattta ccttgattga aataaacgaa
     6841 ataaaaactc gcagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa
     6901 acagcctcta caaataattt tgtttaaata ggtcccaata taaggagagg gtgtacaatg
     6961 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     7021 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
     7081 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
     7141 actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat gaaacagcat
     7201 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa
     7261 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     7321 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     7381 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     7441 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
     7501 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     7561 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
     7621 gagtttgtaa cagctgctgg gattacacat ggcatggatg atctctacaa ataagcttgg
     7681 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     7741 ttctgaggtc attactggat ctatcaacag gagtccaagc g
//
